NL4-3 mCherry Luciferase

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载体基本信息

质粒名称: NL4-3 mCherry Luciferase 
出品公司: 爱迪基因
目录编号: 44965
存储实验室: Warner Greene

载体骨架基本信息
载体名称: pNLENG1
载体出品公司: AIDS Research and Reference Reagent Program
载体抗性: 氨苄青霉素
空载体大小: --
完整质粒大小: 12424 bp
载体修饰: --
载体类型: 慢病毒载体
筛选标记: mCherry, Luciferase
高拷贝/低拷贝: --
生长条件: 30 ℃

插入基因信息
插入基因名称: mCherry, Luciferase, IRES,HIV 1 reporter virus
物种来源: Lentivirus, Human specific
插入大小: --
基因突变: S34C mutation in Nef
GenBank编号 --
启动子 EF-1 alpha– driven mCherry:Puromycin
标签/融合蛋白:
带D212G的mCherry(插接件上的C端子)
荧光素酶,IRES(C端子插入)
克隆方法: 限制性内切酶
5' 克隆位点: BamHI (未破坏)
3' 克隆位点: XhoI(在克隆过程中被破坏)
5' 测序引物 from pNLENG1 5'
3' 测序引物 from pNLENG1 3'

文献引用方法
材料和方法部分: NL4-3 mCherry Luciferase was a gift from Warner Greene (Addgene plasmid # 44965 ; http://n2t.net/addgene:44965 ; RRID:Addgene_44965)
参考文献部分: A flexible model of HIV-1 latency permitting evaluation of many primary CD4 T-cell reservoirs. Lassen KG, Hebbeler AM, Bhattacharyya D, Lobritz MA, Greene WC. PLoS One. 2012;7(1):e30176. doi: 10.1371/journal.pone.0030176. Epub 2012 Jan 24. 10.1371/journal.pone.0030176 PubMed 22291913

载体质粒图谱和多克隆位点信息

NL4-3 mCherry Luciferase质粒图谱

载体简介

载体序列

LOCUS       NL4-3 mCherry Luciferase            17833 bp ds-DNA     circular SYN 16-DEC-2019
DEFINITION  HIV dual reporter vector expressing mCh.
ACCESSION   .
VERSION     .
KEYWORDS    NL4-3 mCherry Luciferase
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17833)
  AUTHORS   Lassen KG, Hebbeler AM, Bhattacharyya D, Lobritz MA, Greene WC
  TITLE     A flexible model of HIV-1 latency permitting evaluation of many 
            primary CD4 T-cell reservoirs.
  JOURNAL   PLoS One. 2012;7(1):e30176. doi: 10.1371/journal.pone.0030176. Epub 
            2012 Jan 24.
  PUBMED    22291913
REFERENCE   2  (bases 1 to 17833)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 16, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..17833
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             377..634
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     CDS             790..2292
                     /codon_start=1
                     /gene="gag"
                     /product="gag protein from human immunodeficiency virus 1"
                     /label=HIV-1 gag
                     /translation="MGARASVLSGGELDKWEKIRLRPGGKKQYKLKHIVWASRELERFA
                     VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTIAVLYCVHQRIDVKDTKEALDKI
                     EEEQNKSKKKAQQAAADTGNNSQVSQNYPIVQNLQGQMVHQAISPRTLNAWVKVVEEKA
                     FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRLHPVHAGP
                     IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
                     ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
                     ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
                     HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
                     PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
     CDS             2085..5096
                     /codon_start=1
                     /gene="pol"
                     /product="pol protein from human immunodeficiency virus 1"
                     /label=HIV-1 pol
                     /translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG
                     ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLDTGADDTVLEEMNLPGRWKPKMIG
                     GIGGFIKVRQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV
                     PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD
                     STKWRKLVDFRELNKRTQDFWEVQLGIPHPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK
                     YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMD
                     DLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI
                     VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL
                     AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT
                     NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP
                     LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ
                     AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH
                     KGIGGNEQVDKLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA
                     SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET
                     AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK
                     ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK
                     QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ
                     MAGDDCVASRQDED"
     misc_feature    4781..4898
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    7759..7992
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8515..8541
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             8787..9494
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEGYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(8932..8950)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9200..9219
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             9501..9554
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9558..11207
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(9621..9639)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    11212..11794
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(11378..11395)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     11605..11624
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             12084..12717
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(14357..14374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14528..15116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14608..14627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(15287..16147)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     15910..15929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16148..16252)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     16320..16338
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(16376..16398)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     16498..16517
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggaagggct aatttggtcc caaaaaagac aagagatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
      121 tgacctttgg atggtgcttc aagttagtac cagttgaacc agagcaagta gaagaggcca
      181 atgaaggaga gaacaacagc ttgttacacc ctatgagcca gcatgggatg gaggacccgg
      241 agggagaagt attagtgtgg aagtttgaca gcctcctagc atttcgtcac atggcccgag
      301 agctgcatcc ggagtactac aaagactgct gacatcgagc tttctacaag ggactttccg
      361 ctggggactt tccagggagg tgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 gctacatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgctca aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagtaaa gccagaggag atctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcggta ttaagcgggg gagaattaga taaatgggaa
      841 aaaattcggt taaggccagg gggaaagaaa caatataaac taaaacatat agtatgggca
      901 agcagggagc tagaacgatt cgcagttaat cctggccttt tagagacatc agaaggctgt
      961 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     1021 ttatataata caatagcagt cctctattgt gtgcatcaaa ggatagatgt aaaagacacc
     1081 aaggaagcct tagataagat agaggaagag caaaacaaaa gtaagaaaaa ggcacagcaa
     1141 gcagcagctg acacaggaaa caacagccag gtcagccaaa attaccctat agtgcagaac
     1201 ctccaggggc aaatggtaca tcaggccata tcacctagaa ctttaaatgc atgggtaaaa
     1261 gtagtagaag agaaggcttt cagcccagaa gtaataccca tgttttcagc attatcagaa
     1321 ggagccaccc cacaagattt aaataccatg ctaaacacag tggggggaca tcaagcagcc
     1381 atgcaaatgt taaaagagac catcaatgag gaagctgcag aatgggatag attgcatcca
     1441 gtgcatgcag ggcctattgc accaggccag atgagagaac caaggggaag tgacatagca
     1501 ggaactacta gtacccttca ggaacaaata ggatggatga cacataatcc acctatccca
     1561 gtaggagaaa tctataaaag atggataatc ctgggattaa ataaaatagt aagaatgtat
     1621 agccctacca gcattctgga cataagacaa ggaccaaagg aaccctttag agactatgta
     1681 gaccgattct ataaaactct aagagccgag caagcttcac aagaggtaaa aaattggatg
     1741 acagaaacct tgttggtcca aaatgcgaac ccagattgta agactatttt aaaagcattg
     1801 ggaccaggag cgacactaga agaaatgatg acagcatgtc agggagtggg gggacccggc
     1861 cataaagcaa gagttttggc tgaagcaatg agccaagtaa caaatccagc taccataatg
     1921 atacagaaag gcaattttag gaaccaaaga aagactgtta agtgtttcaa ttgtggcaaa
     1981 gaagggcaca tagccaaaaa ttgcagggcc cctaggaaaa agggctgttg gaaatgtgga
     2041 aaggaaggac accaaatgaa agattgtact gagagacagg ctaatttttt agggaagatc
     2101 tggccttccc acaagggaag gccagggaat tttcttcaga gcagaccaga gccaacagcc
     2161 ccaccagaag agagcttcag gtttggggaa gagacaacaa ctccctctca gaagcaggag
     2221 ccgatagaca aggaactgta tcctttagct tccctcagat cactctttgg cagcgacccc
     2281 tcgtcacaat aaagataggg gggcaattaa aggaagctct attagataca ggagcagatg
     2341 atacagtatt agaagaaatg aatttgccag gaagatggaa accaaaaatg atagggggaa
     2401 ttggaggttt tatcaaagta agacagtatg atcagatact catagaaatc tgcggacata
     2461 aagctatagg tacagtatta gtaggaccta cacctgtcaa cataattgga agaaatctgt
     2521 tgactcagat tggctgcact ttaaattttc ccattagtcc tattgagact gtaccagtaa
     2581 aattaaagcc aggaatggat ggcccaaaag ttaaacaatg gccattgaca gaagaaaaaa
     2641 taaaagcatt agtagaaatt tgtacagaaa tggaaaagga aggaaaaatt tcaaaaattg
     2701 ggcctgaaaa tccatacaat actccagtat ttgccataaa gaaaaaagac agtactaaat
     2761 ggagaaaatt agtagatttc agagaactta ataagagaac tcaagatttc tgggaagttc
     2821 aattaggaat accacatcct gcagggttaa aacagaaaaa atcagtaaca gtactggatg
     2881 tgggcgatgc atatttttca gttcccttag ataaagactt caggaagtat actgcattta
     2941 ccatacctag tataaacaat gagacaccag ggattagata tcagtacaat gtgcttccac
     3001 agggatggaa aggatcacca gcaatattcc agtgtagcat gacaaaaatc ttagagcctt
     3061 ttagaaaaca aaatccagac atagtcatct atcaatacat ggatgatttg tatgtaggat
     3121 ctgacttaga aatagggcag catagaacaa aaatagagga actgagacaa catctgttga
     3181 ggtggggatt taccacacca gacaaaaaac atcagaaaga acctccattc ctttggatgg
     3241 gttatgaact ccatcctgat aaatggacag tacagcctat agtgctgcca gaaaaggaca
     3301 gctggactgt caatgacata cagaaattag tgggaaaatt gaattgggca agtcagattt
     3361 atgcagggat taaagtaagg caattatgta aacttcttag gggaaccaaa gcactaacag
     3421 aagtagtacc actaacagaa gaagcagagc tagaactggc agaaaacagg gagattctaa
     3481 aagaaccggt acatggagtg tattatgacc catcaaaaga cttaatagca gaaatacaga
     3541 agcaggggca aggccaatgg acatatcaaa tttatcaaga gccatttaaa aatctgaaaa
     3601 caggaaagta tgcaagaatg aagggtgccc acactaatga tgtgaaacaa ttaacagagg
     3661 cagtacaaaa aatagccaca gaaagcatag taatatgggg aaagactcct aaatttaaat
     3721 tacccataca aaaggaaaca tgggaagcat ggtggacaga gtattggcaa gccacctgga
     3781 ttcctgagtg ggagtttgtc aatacccctc ccttagtgaa gttatggtac cagttagaga
     3841 aagaacccat aataggagca gaaactttct atgtagatgg ggcagccaat agggaaacta
     3901 aattaggaaa agcaggatat gtaactgaca gaggaagaca aaaagttgtc cccctaacgg
     3961 acacaacaaa tcagaagact gagttacaag caattcatct agctttgcag gattcgggat
     4021 tagaagtaaa catagtgaca gactcacaat atgcattggg aatcattcaa gcacaaccag
     4081 ataagagtga atcagagtta gtcagtcaaa taatagagca gttaataaaa aaggaaaaag
     4141 tctacctggc atgggtacca gcacacaaag gaattggagg aaatgaacaa gtagataaat
     4201 tggtcagtgc tggaatcagg aaagtactat ttttagatgg aatagataag gcccaagaag
     4261 aacatgagaa atatcacagt aattggagag caatggctag tgattttaac ctaccacctg
     4321 tagtagcaaa agaaatagta gccagctgtg ataaatgtca gctaaaaggg gaagccatgc
     4381 atggacaagt agactgtagc ccaggaatat ggcagctaga ttgtacacat ttagaaggaa
     4441 aagttatctt ggtagcagtt catgtagcca gtggatatat agaagcagaa gtaattccag
     4501 cagagacagg gcaagaaaca gcatacttcc tcttaaaatt agcaggaaga tggccagtaa
     4561 aaacagtaca tacagacaat ggcagcaatt tcaccagtac tacagttaag gccgcctgtt
     4621 ggtgggcggg gatcaagcag gaatttggca ttccctacaa tccccaaagt caaggagtaa
     4681 tagaatctat gaataaagaa ttaaagaaaa ttataggaca ggtaagagat caggctgaac
     4741 atcttaagac agcagtacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     4801 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     4861 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     4921 gagatccagt ttggaaagga ccagcaaagc tcctctggaa aggtgaaggg gcagtagtaa
     4981 tacaagataa tagtgacata aaagtagtgc caagaagaaa agcaaagatc atcagggatt
     5041 atggaaaaca gatggcaggt gatgattgtg tggcaagtag acaggatgag gattaacaca
     5101 tggaaaagat tagtaaaaca ccatatgtat atttcaagga aagctaagga ctggttttat
     5161 agacatcact atgaaagtac taatccaaaa ataagttcag aagtacacat cccactaggg
     5221 gatgctaaat tagtaataac aacatattgg ggtctgcata caggagaaag agactggcat
     5281 ttgggtcagg gagtctccat agaatggagg aaaaagagat atagcacaca agtagaccct
     5341 gacctagcag accaactaat tcatctgcac tattttgatt gtttttcaga atctgctata
     5401 agaaatacca tattaggacg tatagttagt cctaggtgtg aatatcaagc aggacataac
     5461 aaggtaggat ctctacagta cttggcacta gcagcattaa taaaaccaaa acagataaag
     5521 ccacctttgc ctagtgttag gaaactgaca gaggacagat ggaacaagcc ccagaagacc
     5581 aagggccaca gagggagcca tacaatgaat ggacactaga gcttttagag gaacttaaga
     5641 gtgaagctgt tagacatttt cctaggatat ggctccataa cttaggacaa catatctatg
     5701 aaacttacgg ggatacttgg gcaggagtgg aagccataat aagaattctg caacaactgc
     5761 tgtttatcca tttcagaatt gggtgtcgac atagcagaat aggcgttact cgacagagga
     5821 gagcaagaaa tggagccagt agatcctaga ctagagccct ggaagcatcc aggaagtcag
     5881 cctaaaactg cttgtaccaa ttgctattgt aaaaagtgtt gctttcattg ccaagtttgt
     5941 ttcatgacaa aagccttagg catctcctat ggcaggaaga agcggagaca gcgacgaaga
     6001 gctcatcaga acagtcagac tcatcaagct tctctatcaa agcagtaagt agtacatgta
     6061 atgcaaccta taatagtagc aatagtagca ttagtagtag caataataat agcaatagtt
     6121 gtgtggtcca tagtaatcat agaatatagg aaaatattaa gacaaagaaa aatagacagg
     6181 ttaattgata gactaataga aagagcagaa gacagtggca atgagagtga aggagaagta
     6241 tcagcacttg tggagatggg ggtggaaatg gggcaccatg ctccttggga tattgatgat
     6301 ctgtagtgct acagaaaaat tgtgggtcac agtctattat ggggtacctg tgtggaagga
     6361 agcaaccacc actctatttt gtgcatcaga tgctaaagca tatgatacag aggtacataa
     6421 tgtttgggcc acacatgcct gtgtacccac agaccccaac ccacaagaag tagtattggt
     6481 aaatgtgaca gaaaatttta acatgtggaa aaatgacatg gtagaacaga tgcatgagga
     6541 tataatcagt ttatgggatc aaagcctaaa gccatgtgta aaattaaccc cactctgtgt
     6601 tagtttaaag tgcactgatt tgaagaatga tactaatacc aatagtagta gcgggagaat
     6661 gataatggag aaaggagaga taaaaaactg ctctttcaat atcagcacaa gcataagaga
     6721 taaggtgcag aaagaatatg cattctttta taaacttgat atagtaccaa tagataatac
     6781 cagctatagg ttgataagtt gtaacacctc agtcattaca caggcctgtc caaaggtatc
     6841 ctttgagcca attcccatac attattgtgc cccggctggt tttgcgattc taaaatgtaa
     6901 taataagacg ttcaatggaa caggaccatg tacaaatgtc agcacagtac aatgtacaca
     6961 tggaatcagg ccagtagtat caactcaact gctgttaaat ggcagtctag cagaagaaga
     7021 tgtagtaatt agatctgcca atttcacaga caatgctaaa accataatag tacagctgaa
     7081 cacatctgta gaaattaatt gtacaagacc caacaacaat acaagaaaaa gtatccgtat
     7141 ccagagggga ccagggagag catttgttac aataggaaaa ataggaaata tgagacaagc
     7201 acattgtaac attagtagag caaaatggaa tgccacttta aaacagatag ctagcaaatt
     7261 aagagaacaa tttggaaata ataaaacaat aatctttaag caatcctcag gaggggaccc
     7321 agaaattgta acgcacagtt ttaattgtgg aggggaattt ttctactgta attcaacaca
     7381 actgtttaat agtacttggt ttaatagtac ttggagtact gaagggtcaa ataacactga
     7441 aggaagtgac acaatcacac tcccatgcag aataaaacaa tttataaaca tgtggcagga
     7501 agtaggaaaa gcaatgtatg cccctcccat cagtggacaa attagatgtt catcaaatat
     7561 tactgggctg ctattaacaa gagatggtgg taataacaac aatgggtccg agatcttcag
     7621 acctggagga ggcgatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     7681 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     7741 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     7801 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctgatatagt
     7861 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     7921 agtctggggc atcaaacagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     7981 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     8041 ttggaatgct agttggagta ataaatctct ggaacagatt tggaataaca tgacctggat
     8101 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     8161 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     8221 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     8281 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     8341 gcagggatat tcaccattat cgtttcagac ccacctccca atcccgaggg gacccgacag
     8401 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     8461 gaacggatcc ttagcactta tctgggacga tctgcggagc ctgtgcctct tcagctacca
     8521 ccgcttgaga gacttactct tgattgtaac gaggattgtg gaacttctgg gacgcagggg
     8581 gtgggaagcc ctcaaatatt ggtggaatct cctacagtat tggagtcagg aactaaagaa
     8641 tagtgctgtt aacttgctca atgccacagc catagcagta gctgagggga cagatagggt
     8701 tatagaagta ttacaagcag cttatagagc tattcgccac atacctagaa gaataagaca
     8761 gggcttggaa aggattttgc tataagatgg tgagcaaggg cgaggaggat aacatggcca
     8821 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     8881 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     8941 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     9001 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     9061 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     9121 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     9181 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     9241 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     9301 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     9361 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     9421 gctacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     9481 acgagctgta caagccccgg gagggcagag gaagtcttct aacatgcggt gacgtggagg
     9541 agaatcccgg ccctcgagaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
     9601 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
     9661 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
     9721 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
     9781 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
     9841 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
     9901 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
     9961 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
    10021 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
    10081 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
    10141 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
    10201 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
    10261 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
    10321 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
    10381 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
    10441 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
    10501 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
    10561 tcaggcaagg atatgggctc actgagacta catcagctat tctgattacg cccgaggggg
    10621 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
    10681 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
    10741 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
    10801 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
    10861 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
    10921 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
    10981 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
    11041 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
    11101 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
    11161 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaagga gcccctctcc
    11221 ctcccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
    11281 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
    11341 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
    11401 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
    11461 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
    11521 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
    11581 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
    11641 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
    11701 acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
    11761 tcctttgaaa aacacgatga taatggccac aaccatgggt ggcaagtggt caaaaagtag
    11821 tgtgattgga tggcctgctg taagggaaag aatgagacga gctgagccag cagcagatgg
    11881 ggtgggagca gtatgtcgag acctagaaaa acatggagca atcacaagta gcaatacagc
    11941 agctaacaat gctgcttgtg cctggctaga agcacaagag gaggaagagg tgggttttcc
    12001 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
    12061 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaaagaa gacaagatat
    12121 ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga actacacacc
    12181 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
    12241 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
    12301 cctgcatgga atggatgacc ctgagagaga agtgttagag tggaggtttg acagccgcct
    12361 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgacatcg
    12421 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
    12481 tggggagtgg cgagccctca gatgctgcat ataagcagct gctttttgcc tgtactgggt
    12541 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
    12601 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
    12661 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaccc
    12721 aggaggtaga ggttgcagtg agccaagatc gcgccactgc attccagcct gggcaagaaa
    12781 acaagactgt ctaaaataat aataataagt taagggtatt aaatatattt atacatggag
    12841 gtcataaaaa tatatatatt tgggctgggc gcagtggctc acacctgcgc ccggcccttt
    12901 gggaggccga ggcaggtgga tcacctgagt ttgggagttc cagaccagcc tgaccaacat
    12961 ggagaaaccc cttctctgtg tatttttagt agattttatt ttatgtgtat tttattcaca
    13021 ggtatttctg gaaaactgaa actgtttttc ctctactctg ataccacaag aatcatcagc
    13081 acagaggaag acttctgtga tcaaatgtgg tgggagaggg aggttttcac cagcacatga
    13141 gcagtcagtt ctgccgcaga ctcggcgggt gtccttcggt tcagttccaa caccgcctgc
    13201 ctggagagag gtcagaccac agggtgaggg ctcagtcccc aagacataaa cacccaagac
    13261 ataaacaccc aacaggtcca ccccgcctgc tgcccaggca gagccgattc accaagacgg
    13321 gaattaggat agagaaagag taagtcacac agagccggct gtgcgggaga acggagttct
    13381 attatgactc aaatcagtct ccccaagcat tcggggatca gagtttttaa ggataactta
    13441 gtgtgtaggg ggccagtgag ttggagatga aagcgtaggg agtcgaaggt gtccttttgc
    13501 gccgagtcag ttcctgggtg ggggccacaa gatcggatga gccagtttat caatccgggg
    13561 gtgccagctg atccatggag tgcagggtct gcaaaatatc tcaagcactg attgatctta
    13621 ggttttacaa tagtgatgtt accccaggaa caatttgggg aaggtcagaa tcttgtagcc
    13681 tgtagctgca tgactcctaa accataattt cttttttgtt tttttttttt tatttttgag
    13741 acagggtctc actctgtcac ctaggctgga gtgcagtggt gcaatcacag ctcactgcag
    13801 cctcaacgtc gtaagctcaa gcgatcctcc cacctcagcc tgcctggtag ctgagactac
    13861 aagcgacgcc ccagttaatt tttgtatttt tggtagaggc agcgttttgc cgtgtggccc
    13921 tggctggtct cgaactcctg ggctcaagtg atccagcctc agcctcccaa agtgctggga
    13981 caaccggggc cagtcactgc acctggccct aaaccataat ttctaatctt ttggctaatt
    14041 tgttagtcct acaaaggcag tctagtcccc aggcaaaaag ggggtttgtt tcgggaaagg
    14101 gctgttactg tctttgtttc aaactataaa ctaagttcct cctaaactta gttcggccta
    14161 cacccaggaa tgaacaagga gagcttggag gttagaagca cgatggaatt ggttaggtca
    14221 gatctctttc actgtctgag ttataatttt gcaatggtgg ttcaaagact gcccgcttct
    14281 gacaccagtc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
    14341 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
    14401 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
    14461 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
    14521 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
    14581 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
    14641 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
    14701 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
    14761 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
    14821 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
    14881 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
    14941 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
    15001 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
    15061 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
    15121 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
    15181 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
    15241 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
    15301 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
    15361 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
    15421 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
    15481 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
    15541 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
    15601 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
    15661 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
    15721 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
    15781 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    15841 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    15901 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    15961 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    16021 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    16081 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    16141 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    16201 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    16261 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
    16321 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
    16381 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
    16441 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
    16501 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
    16561 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
    16621 ggcgatcggt gcgggcctct tcgctattac gccaggggag gcagagattg cagtaagctg
    16681 agatcgcagc actgcactcc agcctgggcg acagagtaag actctgtctc aaaaataaaa
    16741 taaataaatc aatcagatat tccaatcttt tcctttattt atttatttat tttctatttt
    16801 ggaaacacag tccttcctta ttccagaatt acacatatat tctatttttc tttatatgct
    16861 ccagtttttt ttagaccttc acctgaaatg tgtgtataca aaatctaggc cagtccagca
    16921 gagcctaaag gtaaaaaata aaataataaa aaataaataa aatctagctc actccttcac
    16981 atcaaaatgg agatacagct gttagcatta aataccaaat aacccatctt gtcctcaata
    17041 attttaagcg cctctctcca ccacatctaa ctcctgtcaa aggcatgtgc cccttccggg
    17101 cgctctgctg tgctgccaac caactggcat gtggactctg cagggtccct aactgccaag
    17161 ccccacagtg tgccctgagg ctgccccttc cttctagcgg ctgcccccac tcggctttgc
    17221 tttccctagt ttcagttact tgcgttcagc caaggtctga aactaggtgc gcacagagcg
    17281 gtaagactgc gagagaaaga gaccagcttt acagggggtt tatcacagtg caccctgaca
    17341 gtcgtcagcc tcacaggggg tttatcacat tgcaccctga cagtcgtcag cctcacaggg
    17401 ggtttatcac agtgcaccct tacaatcatt ccatttgatt cacaattttt ttagtctcta
    17461 ctgtgcctaa cttgtaagtt aaatttgatc agaggtgtgt tcccagaggg gaaaacagta
    17521 tatacagggt tcagtactat cgcatttcag gcctccacct gggtcttgga atgtgtcccc
    17581 cgaggggtga tgactacctc agttggatct ccacaggtca cagtgacaca agataaccaa
    17641 gacacctccc aaggctacca caatgggccg ccctccacgt gcacatggcc ggaggaactg
    17701 ccatgtcgga ggtgcaagca cacctgcgca tcagagtcct tggtgtggag ggagggacca
    17761 gcgcagcttc cagccatcca cctgatgaac agaacctagg gaaagcccca gttctactta
    17821 caccaggaaa ggc
//

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